Squash hybrid ZGNEHH6009 and parents thereof

ABSTRACT

The invention provides seed and plants of squash hybrid ZGNEHH6009 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of squash hybrid ZGNEHH6009 and the parent lines thereof, and to methods for producing a squash plant produced by crossing such plants with themselves or with another squash plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the fruit and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No. 61/649,166, filed May 18, 2012, the entire disclosure of which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, more specifically, to the development of squash hybrid ZGNEHH6009 and the inbred squash lines ZGN-130-1096 and ZGN 130-1041T.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.

Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a squash plant of the hybrid designated ZGNEHH6009, the squash line ZGN-130-1096 or squash line ZGN 130-1041T. Also provided are squash plants having all the physiological and morphological characteristics of such a plant. Parts of these squash plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.

In another aspect of the invention, a plant of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.

The invention also concerns the seed of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T. The squash seed of the invention may be provided as an essentially homogeneous population of squash seed of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of squash plants designated ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T.

In yet another aspect of the invention, a tissue culture of regenerable cells of a squash plant of hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T is provided. The tissue culture will preferably be capable of regenerating squash plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides squash plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T.

In still yet another aspect of the invention, processes are provided for producing squash seeds, plants and fruit, which processes generally comprise crossing a first parent squash plant with a second parent squash plant, wherein at least one of the first or second parent squash plants is a plant of squash line ZGN-130-1096 or squash line ZGN 130-1041T. These processes may be further exemplified as processes for preparing hybrid squash seed or plants, wherein a first squash plant is crossed with a second squash plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of squash line ZGN-130-1096 or squash line ZGN 130-1041T. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent squash plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first and second parent squash plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent squash plants. Yet another step comprises harvesting the seeds from at least one of the parent squash plants. The harvested seed can be grown to produce a squash plant or hybrid squash plant.

The present invention also provides the squash seeds and plants produced by a process that comprises crossing a first parent squash plant with a second parent squash plant, wherein at least one of the first or second parent squash plants is a plant of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T. In one embodiment of the invention, squash seed and plants produced by the process are first generation (F₁) hybrid squash seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F₁ hybrid squash plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F₁ hybrid squash plant and seed thereof.

In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T, wherein said preparing comprises crossing a plant of the hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T. The plant derived from hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.

In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T, wherein the plant has been cultivated to maturity, and (b) collecting at least one squash from the plant.

In still yet another aspect of the invention, the genetic complement of squash hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a squash plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides squash plant cells that have a genetic complement in accordance with the squash plant cells disclosed herein, and seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybrid genetic complements, as represented by squash plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a squash plant of the invention with a haploid genetic complement of a second squash plant, preferably, another, distinct squash plant. In another aspect, the present invention provides a squash plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.

Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.

The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.

Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants, seeds and derivatives of squash hybrid ZGNEHH6009, squash line ZGN-130-1096 and squash line ZGN 130-1041T.

Squash hybrid ZGNEHH6009, also known as SV6009YG, is a multi-virus resistant dark green zucchini hybrid that exhibits a very high yield potential compared to other similar products. The erect, open growth habit improves ease of harvest. Highly uniform and cylindrical fruit shape results in a high proportion of top quality fruit.

ZGN 130-1041T is a green zucchini breeding line with exceptional levels of virus resistance. This genotype in particular shows high level resistance to Cucumber Mosaic Virus (CMV), zucchini yellow mosaic potyvirus (ZYMV), and watermelon mosaic virus (WMV). In addition to the disease resistance, this genotype has a medium green fruit color preferred by growers in many regions, as well as acceptable horticultural characteristics.

A. Origin and Breeding History of Squash Hybrid Zgnehh6009

The parents of hybrid ZGNEHH6009 are ZGN-130-1096 and ZGN 130-1041T.

These parents were created as follows:

Squash line ZGN 130-1041T, also known as ZGN-130-1041T, was developed by a selection strategy involving two components: (1) “CAS 46-2002T,” a breeding line having the characteristics of strong female flower production potential, the ZWC multi-virus resistance transgene, and field tolerance to powdery mildew caused by Sphaerotheca fuliginella (Schlechtend.:Fr.) Pollacci, and (2) “ZGN 47-314,” a proprietary breeding line contributing vigor, dark green fruit color, and a long cylindrical fruit shape. The initial cross was made in Florida in April of 2001.

The F1 generation was self pollinated in a California greenhouse in the fall of 2001, to maximize production of F2 seed. The F2 generation was greenhouse grown in 2002, where approximately 500 seedlings were mechanically inoculated with CMV and ZYMV. After development of symptoms, resistant plants were selected and transplanted to a field, where 42 of the survivors were selected for self pollination based upon fruit type and horticultural characteristics. The F3 generation was grown in Florida during the fall of 2002, where 32 F4 selections were made from 14 of the F3 families. Following self pollination, selections were evaluated for resistance to powdery mildew. For the F4 generation, 12 seeds per selection were grown in the spring of 2003. These seedlings were mechanically inoculated with CMV and ZYMV, and survivors were selected for self pollination based on fruit type and horticultural characteristics. The F5 generation was selected in Florida without inoculation during the fall of 2003, and 4 families were retained for further evaluation.

Beginning with the F6 generation, evaluation of these breeding lines was based on performance of both the breeding lines and hybrids created using these breeding lines. For the line designated ZGN 130-1041T, the F6 generation was self pollinated in a California greenhouse during the winter, concurrently with the production of test hybrids. During the subsequent 4 years, more than 100 genetically unique hybrids have been created and evaluated using this lineage. Evaluations of these hybrids have been performed in at least 8 states in the U.S. Hybrids using ZGN 130-1041T show good horticultural characteristics, field tolerance to powdery mildew caused by Sphaerotheca fuliginella (Schlechtend.:Fr.) Pollacci, and resistance to many strains of ZYMV, WMV, and CMV.

Two additional generations of advancement were completed in 2004 through self pollination of greenhouse grown plants. These additional generations further establish uniformity of the parental line, and the founding stock for ZGN 130-1041T was based on a bulk of seed from three F9 generation plants with the same F8 generation source. The pedigree and lineage of this F9 generation is (CAS 46-2002T X ZGN 47-314)-M-5-4-1-2-1-1 M.

The male parent ZGN-130-1096, also know as ZGN 130-1096, was developed from the following initial germplasm sources:

-   -   Commercial variety “Tigress”     -   Commercial variety “Raven”     -   Italian landrace “Striato di Napoli”     -   Germplasm accession “A98S85” from Universidad de Sonora,         Hermosillo courtesy of Sergio Garza     -   Proprietary inbred line “G710,” originally sourced from the         Royal Sluis vegetable seeds company

ZGN-130-1096, or a similar parent line, can be re-created in as few as 21 generations of selective breeding through the following breeding scheme which was used in its development:

-   -   Generation 1: Create a hybrid between germplasm accession A98S85         and inbred line G710     -   Generation 2: Backcross this hybrid to inbred line G710     -   Generation 3: Self pollinate the hybrid Tigress and also a         selection from the first backcross generation (A98S85/G710*1)     -   Generation 4: Self pollinate the hybrid Raven. Self pollinate F2         generation selections of Tigress and A98S85/G710*1     -   Generations 5-7: Self pollinate selected individuals from all         three lineages (Raven, Tigress, and A98S85/G710*1)     -   Generation 8: Hybridize an F6 generation selection of Tigress to         an F4 generation selection of A98S85/G710*1, self pollinate an         F5 generation selection of Raven.     -   Generation 9: Hybridize the F6 generation selection of Raven to         Striato di     -   Napoli, self pollinate the F1 generation of         TigressF6/(A98S85/G710*1)     -   Generations 10-11: Select and self pollinate individual         selections from both lineages (RavenF6/Striato di Napoli and         TigressF6/(A98S85/G710*1))     -   Generation 12: Hybridize and F3 generation selection of         RavenF6/Striato di Napoli to an F4 generation selection of         TigressF6/(A98S85/G710*1).     -   Generations 13-21: for 9 successive generations, self pollinate         individual plant selections from the cross made in         generation 12. Self pollinated seed from a single F9 generation         plant from this cross was coded as ZGN-130-1096, and subsequent         generations of seed increase were not subject to individual         plant selection. Performance testing of hybrids using this         lineage commenced in Generation 18.

During the crossing and inbreeding generations used to develop this parent line, plants were selected based on various criteria, including resistance to diseases (including Zucchini Yellow Mosaic Virus and Watermelon Mosaic Virus), plant architecture, fruit shape, fruit color, blossom scar, synchronicity of male and female flower production, ease of harvest, blossom scar size, speed of plant growth and development, subjective evaluations of fruit productivity, and adaptation to specific environments (environments included field and/or greenhouse locations in California, Mexico, Guatemala, France, and Florida). After generation 18, selection criteria included hybrid performance.

The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.

B. Physiological and Morphological Characteristics of Squash Hybrid ZGNEHH6009, SQUASH LINE ZGN-130-1096 AND SQUASH LINE ZGN 130-1041T

In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of squash hybrid ZGNEHH6009 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-3.

TABLE 1 Physiological and Morphological Characteristics of Hybrid ZGNEHH6009 Comparison Variety CHARACTERISTIC ZGNEHH6009 Payroll 1. Species Pepo Pepo 2. Kind/Use squash squash 3. Type summer (vegetable marrow) summer 4. Cotyledon length 47.2 mm 44.27 mm width 29.59 mm 29.13 mm apex rounded rounded veining obscure plainly visible color medium green dark green color (RHS Color Chart) 138A 147A Seedling shape of cotyledons elliptic (Cora, Tivoli) elliptic intensity of green color of medium (Cora) dark cotyledons cross section of convex (Bianchini, Yellow straight cotyledons Crookneck) 5. Mature Plant growth habit bush bush plant type prickly prickly 6. Main Stem cross-section shape round round diameter at mid-point of 27.6 mm 29.79 mm 1^(st) internode average length 31.9 cm 38.6 cm average number of 18.55 28.56 internodes Stem color completely green (Becky) completely green intensity of green color medium (Cinderella) dark mottling absent (Cinderella) present tendrils absent to rudimentary absent to rudimentary (Goldrush, Sylvana) Plant growth habit bush (Greyzini) bush branching absent (Goldi) absent bush varieties only: erect to semi-erect (Sardane) semi-erect to horizontal attitude of petiole (excluding lower external leaves) 7. Leaves blade shape refuse ovate blade form deep lobed deep lobed margin dentate dentate margin edges frilled frilled average width 29.2 cm 32.92 cm average length 25.16 cm 28.21 cm leaf surface smooth smooth dorsal surface pubescence soft hairy bristled vental surface pubescence soft hairy bristled color medium green medium green color (RHS Color Chart) 137A 137A leaf blotching blotched with gray blotched with gray leaf blade: size large (Kriti) large leaf blade: incisions deep (Civac) medium leaf blade: intensity of medium (Cora) medium green color of upper surface leaf blade: silvery patches present (Civac) present leaf blade: relative area medium (Ambassador) medium covered by silvery patches average petiole length 33.73 cm 33.15 cm petiole length medium (Goldi) long petiole: number of many (White Bush Scallop) many prickles 8. Flower pistillate flower: average 15.66 cm 16.39 cm diameter pistillate flower: ovary turbinate drum-like pistillate flower: average 2.16 cm 2.14 cm pedicel length pistillate flower: margin curved curved shape pistillate flower: margin frilled frilled edges pistillate flower: average 1.35 mm 1.47 mm sepal width pistillate flower: average 5.56 mm 6.14 mm sepal length pistillate flower: color orange deep yellow pistillate flower: color 28B 17A (RHS Color Chart) staminate flower: average 13.95 mm 19.51 mm sepal length staminate flower: average 1.9 mm 2.75 mm sepal width staminate flower: average 96.31 mm 162.53 mm pedicel length staminate flower: color orange orange female flower: ring at absent (Cinderella, Greyzini) present inner side of corolla male flower: ring at inner present (Goldi) present side of corolla male flower: color of ring green (Austral, Belor, Goldi) yellow and green at inner side of corolla male flower: intensity of medium (Verdi) medium green color of ring at inner side of corolla staminate flower: color 28A N25D 9. Fruit market maturity: average 16.3 cm 15.7 cm length market maturity: average 2.79 cm 2.53 cm width - stem end at market maturity: 2.29 cm 1.56 cm average width - blossom end market maturity: average 166.85 gm 133.2 gm weight market maturity: shape straightneck straightneck according to variety type market maturity: apex rounded rounded market maturity: base rounded rounded market maturity: ribs none inconspicuous market maturity: fruit smooth smooth surface market maturity: warts none none market maturity: blossom slightly extended slightly extended scar button young fruit: ratio length/ large (Carlotta) large maximum diameter (zucchini type varieties) young fruit: general shape cylindrical (Ambassador, cylindrical (zucchini and rounded Ibis) zucchini type varieties) young fruit: main color of green (Elite, Opal, Romano) green skin (excluding color of ribs or grooves) young fruit: intensity of medium (Baccara) medium green color of skin (excluding color of ribs or grooves; only varieties with green color of skin) general shape cylindrical cylindrical length (zucchini type very long (Altea) medium varieties) maximum diameter medium (Opal) medium (zucchini type varieties) ratio length/maximum very large (Tarquinio) large diameter (zucchini type varieties) blossom end (zucchini and rounded rounded neck type varieties) grooves absent present ribs present present protrusion of ribs very weak (Leda, Tivoli) weak main color of skin green (Ambassador, Baby green (excluding color of dots, Bear) patches, stripes and bands) intensity of green color of very dark (Baby Bear, medium skin (only varieties with Sardane) green color of skin) color of ribs compared to same (Grey Zucchini) main color of skin dots present (Gold Rush, Table absent Queen) size of main dots small (Ambassador) secondary green color absent (Grey Zucchini, absent between ribs (excluding Small Sugar) dots) warts on skin absent absent size of flower scar medium (Spidi) very small length of peduncle medium (Cinderella) long color of peduncle green (Ambassador) green intensity of green color of medium (Sunburst) medium peduncle mottling of peduncle absent (Sunburst) absent ripe fruit: main color of green green skin (excluding color of mottles, patches, stripes and bands) ripe fruit: intensity of medium main color of skin (only yellow and orange) ripe fruit: secondary color orange of skin (excluding color of mottles, patches, stripes and bands) ripe fruit: color of flesh yellow (Sunburst, Vegetable yellow Spaghetti) ripe fruit: lignified rind present (Elite, Little Gem, present Scallopini, Yellow Summer Crookneck) ripe fruit: structure of fibrous (Vegetable fibrous flesh Spaghetti) 10. Rind average thickness at 2.9 mm 2.03 mm medial toughness hard hard overall color pattern irregular regular main or ground color green greenish gray main or ground color N189A N189A (RHS Color Chart) 11. Flesh average blossom end 43.2 mm 32.4 mm thickness average medial thickness 42.5 mm 38.1 mm average stem end 40.4 mm 40.1 mm thickness texture (fine, granular, stringy stringy lumpy or stringy) texture (soft, firm or firm firm brittle) texture (dry, moist or moist juicy juicy) flavor insipid insipid quality good good color creamy yellow white color (RHS Color Chart) 2D 155A 12. Seed Cavity average length 33.4 cm 34.37 cm average width 5.95 cm 4.92 cm location conforms to fruit shape conforms to fruit shape placental tissue moderately abundant abundant center core inconspicuous prominent 13. Fruit Stalks average length 2.93 cm 2.68 cm average diameter 2.42 cm 1.85 cm cross-section shape irregular irregular twisting not twisted not twisted tapering tapered not tapered straightness slightly curved straight texture spongy spongy furrows deep deep surface rough rough attachment end slightly expanded slightly expanded detaches with difficulty easily color medium green medium green color (RHS Color Chart) 141A 144A 14. Seeds average length 13.6 mm 14.1 mm average width 7.65 mm 8.32 mm average thickness 2.6 mm 2.42 mm face surface smooth smooth color white cream color (RHS Color Chart) 158C 162D luster dull dull margin curved curved margin edge rounded rounded separation from pulp moderately easy easy average grams per 100 10 gm 11.25 gm seeds average number of seeds 201 180.25 per fruit seed coat normal normal size very small (Jack Be Little) medium shape broad elliptic (Baby Boo) elliptic hull present (Baby Bear, Elite) present appearance of hull fully developed (Elite) rudimentary color of hull whitish (Table Queen) cream fruit type: zucchini fruit: patches, stripes or absent (Ambassador, Black bands in ripe stage (if Jack) zucchini type) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of Line ZGN-130-1096 Comparison Variety CHARACTERISTIC ZGN-130-1096 Payroll 1. Species Pepo Pepo 2. Kind/Use squash squash 3. Type summer (vegetable marrow) summer 4. Cotyledon length 43.5 mm 50.1 mm width 31.4 mm 30.35 mm apex tapered rounded veining plainly visible plainly visible color medium green dark green color (RHS Color Chart) 137C 147A Seedling shape of cotyledons elliptic (Cora, Tivoli) elliptic intensity of green color of medium (Cora) dark cotyledons cross section of concave straight cotyledons 5. Mature Plant growth habit bush bush plant type pilose prickly 6. Main Stem cross-section shape round round diameter at mid-point of 25.6 mm 20.1 mm 1^(st) internode average length 25.9 cm 25.1 cm average number of 23.8 25.4 internodes Stem color completely green (Becky) completely green intensity of green color very dark (Goldrush) dark mottling absent (Cinderella) present tendrils absent to rudimentary well developed (Goldrush, Sylvana) Plant growth habit bush (Greyzini) bush branching absent (Goldi) absent bush varieties only: erect (Blanche non coureuse semi-erect attitude of petiole 3, Yellow Crookneck) (excluding lower external leaves) 7. Leaves blade shape reniform reniform blade form deep lobed deep lobed margin dentate dentate margin edges frilled frilled average width 28.7 cm 34.5 cm average length 25.9 cm 29.5 cm leaf surface blistered smooth dorsal surface pubescence glabrous soft hairy vental surface pubescence glabrous soft hairy color dark green medium green color (RHS Color Chart) 137A 137B leaf blotching blotched with gray blotched with gray leaf blade: size medium (Ambassador) medium leaf blade: incisions deep (Civac) medium leaf blade: intensity of dark (Everest) medium green color of upper surface leaf blade: silvery patches present (Civac) present leaf blade: relative area small (Aziz) large covered by silvery patches average petiole length 36.45 cm 35.45 cm petiole length medium (Goldi) medium petiole: number of few (Opaline) many prickles 8. Flower pistillate flower: average 9.05 cm 14.4 cm diameter pistillate flower: ovary drum-like drum-like pistillate flower: average 2.3 cm 4.2 cm pedicel length pistillate flower: margin straight curved shape pistillate flower: margin frilled frilled edges pistillate flower: average 1.15 mm 1.2 mm sepal width pistillate flower: average 3.6 mm 6 mm sepal length pistillate flower: color orange deep yellow pistillate flower: color 21A 23A (RHS Color Chart) staminate flower: average 12.3 mm 16.5 mm sepal length staminate flower: average 2 mm 2.3 mm sepal width staminate flower: average 71.1 mm 178.25 mm pedicel length staminate flower: color orange deep yellow female flower: ring at present (Aurore) present inner side of corolla female flower: color of green (Aurore, Early White yellow and green ring at inner side of Bush Scallop, President) corolla female flower: intensity of medium (Samba, Senator) weak green color of ring at inner side of corolla (varieties with green ring at inner side of corolla) male flower: ring at inner present (Goldi) present side of corolla male flower: color of ring green (Austral, Belor, Goldi) green at inner side of corolla male flower: intensity of medium (Verdi) medium green color of ring at inner side of corolla staminate flower: color 17AS 14A 9. Fruit market maturity: average 16.05 cm 18.3 cm length market maturity: average 4 cm 3.9 cm width - stem end at market maturity: 3.65 cm 3.4 cm average width - blossom end market maturity: average 173.5 gm 240.9 gm weight market maturity: shape straightneck straightneck according to variety type market maturity: apex rounded rounded market maturity: base rounded flattened market maturity: ribs inconspicuous inconspicuous market maturity: rib shallow shallow furrow depth market maturity: rib narrow narrow furrow width market maturity: fruit shallowly wavy smooth surface market maturity: warts none none market maturity: blossom slightly extended raised acorn scar button young fruit: ratio length/ large (Carlotta) medium maximum diameter (zucchini type varieties) young fruit: general shape cylindrical (Ambassador, cylindrical (zucchini and rounded Ibis) zucchini type varieties) young fruit: main color of green (Elite, Opal, Romano) green skin (excluding color of ribs or grooves) young fruit: intensity of very dark (Carnaval, dark green color of skin Corsair) (excluding color of ribs or grooves; only varieties with green color of skin) general shape cylindrical club shaped length (zucchini type short (Jedida) short varieties) maximum diameter small (Goldi) small (zucchini type varieties) ratio length/maximum medium (Cora) medium diameter (zucchini type varieties) blossom end (zucchini and rounded rounded neck type varieties) grooves absent present ribs present present protrusion of ribs weak (Ambassador) weak main color of skin green (Ambassador, Baby green (excluding color of dots, Bear) patches, stripes and bands) intensity of green color of very dark (Baby Bear, medium skin (only varieties with Sardane) green color of skin) color of ribs compared to same (Grey Zucchini) same main color of skin dots present (Gold Rush, Table present Queen) size of main dots very small (Badger Cross) very small secondary green color absent (Grey Zucchini, absent between ribs (excluding Small Sugar) dots) warts on skin absent absent size of flower scar small (Goldi) medium length of peduncle long (Tivoli) medium color of peduncle green (Ambassador) green intensity of green color of dark (Gold Rush) medium peduncle mottling of peduncle present (Elite) present ripe fruit: secondary color orange (Autumn Gold) yellow of skin (excluding color of mottles, patches, stripes and bands) ripe fruit: intensity of medium medium main color of skin (only yellow and orange) ripe fruit: secondary color green green of skin ripe fruit: color of flesh yellow (Sunburst, Vegetable cream Spaghetti) ripe fruit: lignified rind present (Elite, Little Gem, present Scallopini, Yellow Summer Crookneck) ripe fruit: structure of fibrous (Vegetable fibrous flesh Spaghetti) 10. Rind average thickness at 2.4 mm 2.1 mm medial toughness hard hard overall color pattern regular regular main or ground color green yellowish green main or ground color N189A 151D (RHS Color Chart) 11. Flesh average blossom end 8.2 mm 9.45 mm thickness average medial thickness 66.7 mm 25.2 mm average stem end 17.45 mm 16.2 mm thickness texture (fine, granular, fine fine lumpy or stringy) texture (soft, firm or firm soft brittle) texture (dry, moist or moist juicy juicy) flavor slightly sweet insipid quality good good color yellowish green creamy white color (RHS Color Chart) 157D 157A 12. Seed Cavity average length 33.15 cm 28.7 cm average width 8.1 cm 5.85 cm location conforms to fruit shape conforms to fruit shape placental tissue abundant abundant center core inconspicuous inconspicuous 13. Fruit Stalks average length 4.05 cm 2.7 cm average diameter 1.8 cm 2.1 cm cross-section shape round irregular twisting twisted not twisted tapering tapered not tapered straightness straight straight texture hard spongy furrows deep deep surface smooth smooth attachment end expanded expanded detaches easily easily color dark green medium green color (RHS Color Chart) 139A 137C 14. Seeds average length 12.2 mm 14.1 mm average width 7.25 mm 8.6 mm average thickness 2.9 mm 1.9 mm face surface wrinkled scaly color white cream color (RHS Color Chart) 155B 162D luster dull dull margin curved curved margin edge rounded rounded separation from pulp moderately easy easy average grams per 100 12.4 gm 11.6 gm seeds average number of seeds 142.5 248.5 per fruit seed coat normal normal size small (Delicata) medium shape elliptic (Elite) broad elliptic hull present (Baby Bear, Elite) present appearance of hull fully developed (Elite) fully developed color of hull cream (De Nice à Fruit cream Rond) fruit type: zucchini fruit: patches, stripes or absent (Ambassador, Black bands in ripe stage (if Jack) zucchini type) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

TABLE 3 Physiological and Morphological Characteristics of Line ZGN 130-1041T CHARACTERISTIC ZGN 130-1041T 1. General Descriptors Fruit Shape/Variety Group Zucchini Expected Primary Usage Culinary Parts of Plant Providing Expected Immature Fruit Primary Usage Cotyledons Measured Between Full Length to Width ratio - 1.75 Expansion of 1^(st) and 2^(nd) True Leaves Apex - Not Notched Veining - Obvious 2. Main Stem Main Color Dark Green (Nearly Entire Length - Fordhook Zucchini, Jack O'Lantern, Howden) White Marks at Nodes Present Yellow Marks (Associated with Absent Precocious Yellow Gene Complex) Growth Habit (20 True Leaves) Bush True-bush (Fordhook Zucchini, Cocozelle, Ronde de Nice, benning's Green Tint) Tendrils (20 True Leaves) Absent or Rudimentary Main Stem Internode Dimensions Observed after 20^(th) Internode Developed Length Constant from 5^(th) to 15^(th) Internode Width Decreases from 5^(th) to 15^(th) Internode 3. Petioles Derived from Main Stem Observed after 20^(th) Node Developed Length to Width Medial Ratio of 49.70 10th Length to Width Medial Ratio of 45.19 15^(th) Petiole Spininess (Prickles) Observed after Moderately Spiny (Cocozelle) 20^(th) Internode Angle of 6^(th) through 15^(th) on Main Horizontal (Caserta, less than 10°) Stem 4. Laminae Lobing of 10^(th) and 15^(th) on Main Medium Lobed Stem Dimensions of Leaf after 20^(th) 0.83 Length to Maximal Width Ratio of 10^(th) True Internode Leaf Dimensions of Leaf after 20^(th) 0.85 Length to Maximal Width ratio of 15^(th) True Internode Leaf Silver Blotching or Mottling Silver Blotching over a Moderate Amount of the Surface 5. Flowers Number per Node One (almost always) (Fordhook Zucchini, Cocozelle) Staminate on day of Anthesis on Main Stem (Between Nodes 11/20) Length from Base of Calyx to 71.6 mm Tip of Corolla Exterior Width at Top of Calyx 17.3 mm Cup Pedicel Length 179.5 mm Length of Anther Column 11.1 mm Dominant Color of Corolla of Orange-yellow (Day of Anthesis) Staminate Flower Ring at Base of Interior of Staminate Absent Corolla Ring at Base of Interior of Pistillate Yellow Corolla Pistillate Flower on Day of Anthesis Length from Base of Calyx to 67.5 mm Tip of Corolla Pedicel Length 16.4 mm Ovary Color (Day Prior to Anthesis) Green (Black Beauty, Fordhook Zucchini, Cocozelle, Clarita) 6. Immature Fruit Size (3-5 Days Past Anthesis) Length (through Axis) to Medial 4.48 Width Ratio Length (through Axis) to 4.01 Maximal Width Ratio Color (3-5 Days Past Anthesis) Intense Green (Fordhook Zucchini, Black Beauty, Jack O'Lantern, Senator, Spineless Beauty, Raven) Fruit Flecks Small (Nero di Milano, Raven, Magic Lantern) Fruit Warting Absent (Cocozelle, Fordhook Zucchini, Ronde de Nice, Gentry) 6. Mature Fruit Dimensions (40 Days Past Anthesis) 3.0 Length (through Axis) to Medial Width Ratio Warting Absent (Cocozelle, Fordhook Zucchini, Ronde de Nice) Stylar Scar Flat Stylar End Nearly Flat (Fordhook Zucchini, True French) Peduncle End Convex Surface Neither Netted or Cracked Exterior Color Black Green (Fordhook Zucchini, Taybelle) 7. Seed Hull (Mature) Present with Normal Appearance *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

C. Breeding Squash Plants

One aspect of the current invention concerns methods for producing seed of squash hybrid ZGNEHH6009 involving crossing squash lines ZGN-130-1096 and ZGN 130-1041T. Alternatively, in other embodiments of the invention, hybrid ZGNEHH6009, line ZGN-130-1096, or line ZGN 130-1041T may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid ZGNEHH6009 and/or the squash lines ZGN-130-1096 and ZGN 130-1041T, or can be used to produce plants that are derived from hybrid ZGNEHH6009 and/or the squash lines ZGN-130-1096 and ZGN 130-1041T. Plants derived from hybrid ZGNEHH6009 and/or the squash lines ZGN-130-1096 and ZGN 130-1041T may be used, in certain embodiments, for the development of new squash varieties.

The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid ZGNEHH6009 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with ZGNEHH6009 and/or squash lines ZGN-130-1096 and ZGN 130-1041T for the purpose of developing novel squash lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of squash plants developed by this invention.

D. Performance Characteristics

As described above, hybrid ZGNEHH6009 exhibits desirable traits, as conferred by squash lines ZGN-130-1096 and ZGN 130-1041T. The performance characteristics of hybrid ZGNEHH6009 and squash lines ZGN-130-1096 and ZGN 130-1041T were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.

TABLE 4 Hybrid performance trial Sept-Oct 2011 at Tifton, GA Comparing ZGNEHH6009 to two leading competitor hybrids, 9 harvest dates. Fruit Fruit Blossom Ease of Growth Marketable Variety Shape Color Uniformity Scar Harvest Vigor Habit Spininess fruit/plant HybridA 4 3 3 4 4 4 4 5 1.62 HybridA 5 5 4 4 3 5 5 4 1.81 HybridB 5 2 3 3 5 3 4 3 1.21 HybridB 4 2 4 4 4 3 4 4 2.37 ZGNEHH6009 4 3 3 5 4 4 4 5 3.90 ZGNEHH6009 3 3 2 6 4 3 4 4 4.79 Hybrid A is variety “Felix” from Harris Moran, Hybrid B is variety “Envy” from Rogers.

TABLE 5 Hybrid performance data, Tifton GA, September-October 2010, 13 harvest dates. Fruit Fruit Blossom Ease of Growth Marketable Hybrid Shape Color Uniformity Scar Harvest Vigor Habit Spininess fruit/plant JUSTICE III 6 6 6 5 1 4 3 5 5.8 JUSTICE III 7 6 7 4 1 4 3 4 6.7 JUSTICE III 7 4 3 5 3 4 4 4 5.7 ZGNEHH6009 4 3 4 6 2 5 3 4 8.1 ZGNEHH6009 6 3 3 7 4 5 5 4 8.1 ZGNEHH6009 4 2 2 5 1 5 5 5 6.9 Traits - Fruit Shape Rating (1-9 scale) where 1 is ideal, 5 is typical commercial hybrid quality, and 9 is unacceptably curved, bulby, or of incorrect length. Fruit Color Rating (1-9 scale) evaluates the evenness of green color on the fruit, where 1 is perfectly uniform color on all fruits, 5 is a commercially average frequency of light fruit discoloration on portions of fruits not exposed to sunlight, and 9 is severe discoloration on the shaded side of the majority of commercial harvest stage fruits. Blossom Scar rating (1-9 scale) 1 is a tiny and inconspicuous blossom scar, likely less than 2 mm in diameter, 5 is a pronounced but commercially acceptable blossom scar averaging in the neighborhood of 4-5 mm, and 9 is a large and/or protruding blossom scar frequently greater than 8 mm. Ease of harvest score (1-9 scale) for hand-harvested fruits (as opposed to knife harvested fruits) - a score of 1 represents 0 broken fruits among the first 9 fruits harvested in a plot, and each score higher represents 1 additional broken fruit among the first 9 fruits harvested. A score of 9 represents 8 or more broken fruits among the first 9 harvested fruits. Vigor score (1-9 scale) subjective rating of plant vigor during the harvest period, 1 represents an extremely vigorous plant, 5 represents a plant of average vigor, and 9 represents an extremely weak plant. Growth Habit score (1-9 scale) a subjective rating of the desirability of the plant type, where 1 represents an ideal plant shape (upright growth, single stem, fruit held above the plant for easy harvest, short internode length), 5 represents a growth habit that is acceptable in many circumstances (somewhat upright or diagonal growth habit, limited branching, most fruit visible when viewed from above), and 9 represents an unacceptable growth habit (completely prostrate growth, long internodes, acute petiole angles that obscure fruit and/or cause damage to fruit upon harvest) Spininess score (1-9 scale), where 1 represents a plant wholly free of spines, 5 represents a moderate level of spininess that can cause moderate damage to fruits at the time of harvest, and 9 represents a high density of large spines that damage fruit and cause discomfort to harvesters. Marketable Fruit/Plant: the number of marketable quality fruits produced in the (10 harvest dates) harvest window.

E. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those squash plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental squash plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental squash plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a squash plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.

In one embodiment, progeny squash plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of squash the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.

New varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.

With the development of molecular markers associated with particular traits, it is possible to add additional traits into an established germ line, such as represented here, with the end result being substantially the same base germplasm with the addition of a new trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for integrating single or multiple traits or QTL into an elite line. This molecular breeding-facilitated movement of a trait or traits into an elite line may encompass incorporation of a particular genomic fragment associated with a particular trait of interest into the elite line by the mechanism of identification of the integrated genomic fragment with the use of flanking or associated marker assays. In the embodiment represented here, one, two, three or four genomic loci, for example, may be integrated into an elite line via this methodology. When this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid. These additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait. In another embodiment, loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to confer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.

Selection of squash plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

F. Plants Derived by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.

An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., BioTechnology, 3(7):637-642, 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S);1 the nopaline synthase promoter (An et al., Plant Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell, 1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem; the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform virus promoter; a commelina yellow mottle virus promoter; and other plant DNA virus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and could potentially be introduced into a squash plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a squash plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.

G. Definitions

In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F₁), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes from different plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenic plants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.

Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a squash variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.

Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.

Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a squash plant by transformation.

H. Deposit Information

A deposit of squash hybrid ZGNEHH6009 and inbred parent lines ZGN-130-1096 and ZGN 130-1041T, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit were Sep. 17, 2012, May 31, 2012, and 30 Apr. 2007, respectively. The accession numbers for those deposited seeds of squash hybrid ZGNEHH6009 and inbred parent lines ZGN-130-1096 and ZGN 130-1041T are ATCC Accession No. PTA-13192, ATCC Accession No. PTA-12929, and ATCC Accession No. PTA-8397, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

All references cited herein are hereby expressly incorporated herein by reference. 

What is claimed is:
 1. A squash plant comprising at least a first set of the chromosomes of squash line ZGN-130-1096, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12929.
 2. A seed comprising at least a first set of the chromosomes of squash line ZGN-130-1096, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12929.
 3. The plant of claim 1, which is inbred.
 4. The plant of claim 1, which is hybrid.
 5. The seed of claim 2, which is inbred.
 6. The seed of claim 2, which is hybrid.
 7. The plant of claim 4, wherein the hybrid plant is squash hybrid ZGNEHH6009, a sample of seed of said hybrid ZGNEHH6009 having been deposited under ATCC Accession Number PTA-13192.
 8. The seed of claim 6, defined as a seed of squash hybrid ZGNEHH6009, a sample of seed of said hybrid ZGNEHH6009 having been deposited under ATCC Accession Number PTA-13192.
 9. The seed of claim 2, defined as a seed of line ZGN-130-1096.
 10. A plant part of the plant of claim
 1. 11. The plant part of claim 10, further defined as a leaf, an ovule, pollen, a fruit, or a cell.
 12. A squash plant having all the physiological and morphological characteristics of the squash plant of claim
 7. 13. A tissue culture of regenerable cells of the plant of claim 1, said cells comprising at least a first set of the chromosomes of squash line ZGN-130-1096, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12929.
 14. The tissue culture according to claim 13, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
 15. A squash plant regenerated from the tissue culture of claim
 13. 16. A method of vegetatively propagating the plant of claim 1, said method comprising the steps of: (a) collecting tissue capable of being propagated from a plant according to claim 1; (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
 17. The method of claim 16, further comprising growing at least a first plant from said rooted plantlets.
 18. A method of introducing a desired trait into a squash line, said method comprising: (a) crossing a plant of line ZGN-130-1096 with a second squash plant that comprises a desired trait to produce F1 progeny, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12929; (b) selecting an F1 progeny that comprises the desired trait; (c) backcrossing the selected F1 progeny with a plant of line ZGN-130-1096 to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristics of squash line ZGN-130-1096 ; and (e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait and otherwise comprises all of the morphological and physiological characteristics of line ZGN-130-1096.
 19. A squash plant produced by the method of claim 18, wherein the plant has the desired trait and otherwise comprises all of the morphological and physiological characteristics of squash line ZGN-130-1096.
 20. A method of producing a plant comprising an added trait, said method comprising introducing a transgene conferring the trait into a plant of hybrid ZGNEHH6009, or line ZGN-130-1096, a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-13192, and ATCC Accession Number PTA-12929, respectively.
 21. A plant produced by the method of claim 20, wherein the plant otherwise comprises all of the morphological and physiological characteristics of squash hybrid ZGNEHH6009 or squash line ZGN-130-1096.
 22. The plant of claim 1, comprising a transgene.
 23. The plant of claim 22, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.
 24. A plant produced by introducing a single locus conversion into a plant of squash line ZGN-130-1096, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12929, wherein said plant comprises the single locus conversion and all of the morphological and physiological characteristics of squash line ZGN-130-1096.
 25. The plant of claim 24, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.
 26. A method for producing a seed of a plant derived from at least one of hybrid ZGNEHH6009, or line ZGN-130-1096, said method comprising the steps of: (a) crossing a squash plant of hybrid ZGNEHH6009, or line ZGN-130-1096 with itself or a second squash plant; a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-13192, and ATCC Accession Number PTA-12929, respectively; and (b) allowing seed of a hybrid ZGNEHH6009, or line ZGN-130-1096-derived squash plant to form.
 27. The method of claim 26, further comprising the steps of: (c) selfing a plant grown from said hybrid ZGNEHH6009, or ZGN-130-1096-derived squash seed to yield additional hybrid ZGNEHH6009, or line ZGN-130-1096-derived squash seed; (d) growing said additional hybrid ZGNEHH6009, or line ZGN-130-1096-derived squash seed of step (c) to yield additional hybrid ZGNEHH6009, or line ZGN-130-1096-derived squash plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid ZGNEHH6009, or line ZGN-130-1096-derived squash plant.
 28. The method of claim 26, wherein the second squash plant is of an inbred squash line.
 29. The method of claim 26, comprising crossing line ZGN-130-1096 with line ZGN 130-1041T, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-12929, and ATCC Accession Number PTA-8397, respectively.
 30. The method of claim 27, further comprising: (f) crossing the further hybrid ZGNEHH6009, or ZGN-130-1096-derived squash plant with a second squash plant to produce seed of a hybrid progeny plant.
 31. A plant part of the plant of claim
 7. 32. The plant part of claim 31, further defined as a leaf, an ovule, pollen, a fruit, or a cell.
 33. A method of producing a squash seed, said method comprising crossing the plant of claim 1 with itself or a second squash plant and allowing seed to form.
 34. A method of producing a squash, said method comprising: (a) obtaining a plant according to claim 1, wherein the plant has been cultivated to maturity; and (b) collecting a squash from the plant.
 35. A method of producing a plant of squash hybrid ZGNEHH6009 further comprising a single locus conversion, said method comprising crossing a plant of line ZGN 130-1041T with a plant of line ZGN-130-1096, wherein at least one of said lines comprises a single locus conversion, a sample of seed of said lines ZGN 130-1041T and ZGN-130-1096 having been deposited under ATCC Accession No. PTA-8397 and ATCC Accession Number PTA-12929, respectively, and wherein said plant comprises all of the morphological and physiological characteristics of squash hybrid ZGNEHH6009. 